Abstract
Purpose: The purpose of this study was to investigate whether infertility is affected by sperm chromatin and cytogenetic abnormalities. To this purpose, the frequency of sperm premature chromosome condensation (PCC) induction and numerical chromosome abnormalities in the sperm of normal and sub-fertile men were analyzed. PCC rate was studied for evaluating the role of sperm chromatin abnormalities in the process of nuclear decondensation.
Design: Controlled prospective study.
Setting: Infertility Genetics Department, Royan Institute.
Patient: Sub-fertile males who were referred for infertility treatment and sperm cytogenetical studies.
Methods: Hamster oocytes were retrieved after super ovulation by PMSG and HCG injection. Following treatment with Hyaloronidase, zona was removed by trypsin digestion. Sperms were classified according to the morphology, movement and counts and then processed by swim up method. After capacitation, zona-free oocytes were incubated with sperms, and then transferred to fresh media containing colcemid. Slides were prepared using Tarkowskie’s standard air-drying technique. Oocytes were analyzed using × 1000 microscope after staining in 5% of Giemsa.
Main outcome measure: The incidence of sperm aneuploidy, PCC and penetration rate in three groups were determined.
Results: Regarding the PCC rate, a significantly higher frequency was found in infertile patients. (P<0.001). The frequency of PCC in oligosperm samples was 36% compared to 19.37% in normal group. A higher frequency of numerical chromosome abnormalities was found in infertile patients. The rate of these abnormalities was 5.6% in normal group and 18.5% in oligospermic samples. Despite the considerable difference between those frequencies, this difference is not significant. (P>0.05)
Conclusions: From the results it can be concluded that, formation of sperm PCC is a major cause of failed fertilization in individuals with sperm abnormalities. PCC may form due to chromatin abnormalities, improper DNA packing, chromosomal abnormalities and penetration delay of sperm. Also this may be involved in the etiology of some cases of idiopathic infertility. About numerical chromosome abnormalities although the differences are not significant, there is an association between sperm numerical chromosome abnormalities and male infertility. These abnormalities can be originated from meiotic process in spermatogenesis.
Keywords: Chromosome abnormalities, hamster zona-free oocyte, human sperm, in-vitro fertilization
Full Text
The Full Text of this article is available as a PDF (115.6 KB).
References
- 1.Martin RH, Rademarker AW. The effect of age on the frequency of sperm chromosomal abnormalities in normal men. Am J Hum Genet. 1987;41:489–492. [PMC free article] [PubMed] [Google Scholar]
- 2.Martin RH: Human sperm chromosome complements; Effect of donor age, freezing and segregation and inversion carriers. Mutation and Environment Part B; 1990; 435–445 [PubMed]
- 3.Testart J, Gautier C. ICSI in infertile patients with structural chromosome abnormalities. Hum Reprod. 1996;11(2):2609–2612. doi: 10.1093/oxfordjournals.humrep.a019179. [DOI] [PubMed] [Google Scholar]
- 4.Colombero LT, June JH. Incidence of sperm aneuploidy in relation to semen characteristics and assisted reproductive outcome. Fertility and Sterility. 1999;72(1):90–96. doi: 10.1016/s0015-0282(99)00158-2. [DOI] [PubMed] [Google Scholar]
- 5.Shi Q, Martin RH. Aneuploidy in human sperm. Cytogenet Cell Genet. 2000;90:219–226. doi: 10.1159/000056773. [DOI] [PubMed] [Google Scholar]
- 6.Mozdarani H, Aghdaei F. Cytogenetic analysis of failed fertilized oocytes from Iranian infertile women after IVF and ICSI procedures. Middle East Fertility Soc J. 2001;6(3):216–225. [Google Scholar]
- 7.Johnson RT, Rao PN. Mammalian cell fusion; Induction of premature chromosome condensation in interphase nuclei. Nature. 1970;226:717–722. doi: 10.1038/226717a0. [DOI] [PubMed] [Google Scholar]
- 8.Schmiday H, Sperling K. Prematurely condensed human sperm chromosomes after (IVF) Hum Genet. 1986;74:441–443. doi: 10.1007/BF00280502. [DOI] [PubMed] [Google Scholar]
- 9.Schmiday H, Kentenich H. PCC after IVF. Hum Reprod. 1989;4(6):989–995. [Google Scholar]
- 10.Schmiday H, Tandler-Schneider A. Premature chromosome condensation of the sperm nucleus after ICSI. Hum Reprod. 1996;11(10):2239–2245. doi: 10.1093/oxfordjournals.humrep.a019083. [DOI] [PubMed] [Google Scholar]
- 11.Rosenbuch BE. Frequency and patterns of premature sperm chromosome condensation in oocytes facility to fertilize after ICIS. J. Assisted Reprod Genet. 2000;17(5):253–259. doi: 10.1023/A:1009454231659. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 12.Masui Y, Markert CL. Cytoplasmic control of nuclear behavior during meiotic maturation of frog oocytes. J Exp Zool. 1971;117:129–146. doi: 10.1002/jez.1401770202. [DOI] [PubMed] [Google Scholar]
- 13.Murragy AW, Kirschner MW. Cyclin synthesis drives the early embryonic cell cycle. Nature. 1989;339:275–280. doi: 10.1038/339275a0. [DOI] [PubMed] [Google Scholar]
- 14.Sakkas D, Urner F. Sperm chromatin anomalies can influence decondensation after ICSI. Hum Reprod. 1996;11(4):837–843. doi: 10.1093/oxfordjournals.humrep.a019263. [DOI] [PubMed] [Google Scholar]
- 15.Tesarik J, Sousa M. Spermatids as gametes: indications and limitations. Hum Reprod. 1998;13(3):89–107. doi: 10.1093/humrep/13.suppl_3.89. [DOI] [PubMed] [Google Scholar]
- 16.Esterhuizen AD, Franken DR. Chromatin packaging as an indicator of human sperm disfunction. J Assist Reprod Genet. 2000;17(9):508–514. doi: 10.1023/A:1009493824953. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 17.Tarkowski AK. An Air-drying method for chromosome preparation from mouse eggs. Cytogenetics. 1986;5:394–400. [Google Scholar]
- 18.Palermo G, Joris H. Sperm characteristics and outcome of human assisted fertilization by sub zonal insemination and ICSI. Fertility and Sterility. 1993;59:826–835. doi: 10.1016/s0015-0282(16)55867-1. [DOI] [PubMed] [Google Scholar]
- 19.Dozortser D, Desutter P. Behavior of spermatozoa in human oocytes displaying no or one pronucleus after ICSI. Hum Reprod. 1994;9:2134–2144. doi: 10.1093/oxfordjournals.humrep.a138406. [DOI] [PubMed] [Google Scholar]
- 20.Bianchi PG, Mani C. Effect of DNA protamination on fluorochrome staining and in-situ nick-translation of murine and human mature spermatozoa. Biol Reprod. 1993;49:1038–1043. doi: 10.1095/biolreprod49.5.1083. [DOI] [PubMed] [Google Scholar]
- 21.Bianchi PG, Mani C. Chromatin packaging and morphology in ejaculated human spermatozoa: evidence of hidden anomalies in normal spermatozoa. Mol Hum Reprod. 1996;2:139–144. doi: 10.1093/molehr/2.3.139. [DOI] [PubMed] [Google Scholar]
- 22.Manicardi GC, Bianchi PG. Underprotamination and nicking of DNA in ejaculated human spermatozoa are highly related phenomena. Biol Reprod. 1995;52:864–867. doi: 10.1095/biolreprod52.4.864. [DOI] [PubMed] [Google Scholar]
- 23.Sailer BI, Jost LK. Mammalian sperm DNA susceptibility to in-situ denaturation associated with the presence of DNA strand breaks as measured by the terminal deoxynucleotidyl transferase assay. J Androl. 1995;16:80–87. [PubMed] [Google Scholar]
- 24.Perreault SD, Naish SJ. The timing of Hamster sperm nuclear decondensation and male pronucleus formation is related to sperm nuclear disulfide bond content. Biol Reprod. 1987;36:239–244. doi: 10.1095/biolreprod36.1.239. [DOI] [PubMed] [Google Scholar]
- 25.Balhorn R, Reed S. Aberrant protamine 1/Protamine 2 ratios in sperm of infertile human males. Experimental. 1988;44:52–55. doi: 10.1007/BF01960243. [DOI] [PubMed] [Google Scholar]
- 26.Belokopytova LA, Kostyleva E. Human male infertility may be due to a decrease of the protamine (P2) content of sperm chromatin. Mal Reprod Deve. 1993;34:53–57. doi: 10.1002/mrd.1080340109. [DOI] [PubMed] [Google Scholar]
- 27.Yebra De, Balleson L. Complete selective absence of protamine P2 in human. J Biol Chem. 1993;268:10553–10557. [PubMed] [Google Scholar]
- 28.Moosani N, Cox M. Chromosomal analysis of sperm from men with idiopathic infertility using sperm karyotyping and FISH. Fertility and Sterility. 1995;64(4):811–817. doi: 10.1016/s0015-0282(16)57859-5. [DOI] [PubMed] [Google Scholar]